Development of structured illumination microscope using transmission
diffraction grating obtained by analogue microfilming method
Abstract
We present the development of a custom-built structured illumination
microscope (SIM) featuring a specially designed transmission diffraction
grating. Employing the analogue microfilming method, we fabricated
transmission diffraction gratings suited to the specific requirements of
our system. This robust and cost-effective method allows for the
fabrication of diffraction gratings with customised constants, ensuring
excellent transmission in both the visible and near-infrared spectra.
Additionally, to assess the performance of our system, we measured the
resolution in both epifluorescent and superresolution imaging modalities
by applying two independent methods: the conventional resolution
measurement using fluorescent beads and the knife-edge technique applied
on the MoS 2 monolayer flakes. Both methods confirmed
enhancement in the resolution of SIM over the epifluorescent imaging
modality. Furthermore, we have successfully demonstrated the
capabilities of our microscope by imaging fluorescently labelled
astrocytes, specifically targeting the vimentin filament protein within
these cells. The superresolution images reveal fine structures of the
vimentin cytoskeleton that remain unresolved in the epifluorescent
image.